Work Package 5 - Genocedar

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Work Package 5

Work Packages
WP5: Application and Analysis of Molecular Markers - Gene Amplification and Nucleotide Diversity

The present study will apply a genetic technology towards the identification of key genes determining adaptive traits in the target species. Genomics is nowadays a powerful molecular tool for directly assessing genes diversity in wild plant species. The proposed research activities aim to increase genomic resources and study candidate genes involved in the regulation of coniferous development, growth and response to environmental stress, by testing / using available adaptive genes from the GenBanks and global literature in Cedrus spp. and/or the Pinacea family.

Initially, the genes which successfully amplified C. brevifolia at an earlier phylogenetic study of Cedrus spp.
(Karam 2010), will be tested in this project. In addition, transferability will be tested for the 60 genes which were recently developed for the congener cedar species (C. libani and C. atlantica). This transferability is expected to be high (e.g. 10 - 50 genes), as a previous study on neutral markers (microsatellites) showed 66.7% transferability (relatively high for conifer species) from C. atlantica to C. brevifolia (Eliades 2008).

The successfully amplified genes will be applied for the whole sample size, for assessing the adaptation and adaptability of C. brevifolia. PCR protocols optimisation may be needed, while cloning of amplified fragments using external vector in agar gel may be performed for clearer nucleotide diversity and complete sequencing of the amplified fragments. Sequence alignment will be done using suitable software, thus leading to nucleotide diversity detection; the three-dimensional protein structure will also be shaped.

Two deliverables (D) are included in this WP:
D13: Principles and Laboratory Protocols of Molecular Techniques Employed in assessments for the present project.
D14: Preliminary report with the statistic measurements of population diversity.

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